Characterization and Genetics of KI toxicity in iPSC-derived cardiomyocytes

摘要 激酶抑制剂（KIs）的发展已经彻底改变了广谱性肿瘤的治疗。 癌症类型。然而，与药物相关的不良反应往往会抵消抗癌的益处 毒品随着KI的广泛使用，很明显，一部分患者 会出现心脏功能障碍和心力衰竭目前我们对潜在的 病理生理学和危险因素是有限的。实验侧重于了解 潜在的心脏毒性的分子和遗传机制将是至关重要的，以改善和指导 进一步的药物开发。人诱导多能干细胞的研究进展 hiPSC（hiPSC）和将hiPSC分化为CM（hiPSC-CM）的能力提供了前所未有的优势。 疾病建模和心脏毒性测试的机会。人类iPSC-CM表现出 性质与其主要对应物高度相似，因此提供了相关的“盘中病人” 模型该提案的总体目标是确定和理解分子和 使用hiPSC-CM研究KI诱导心脏毒性的遗传机制。 该提案建立在通过各种NHLBI赠款资助的广泛基础设施之上。我们有 产生了250个hiPSC系，并将这些系分化成来自参与者的CM。 NHLBI HyperGEN队列。我们建议使用这些hiPSC-CM来识别潜在的通路 和遗传标记，这有助于对KI的反应的变异性。使用表达式 分析，我们将描述与暴露hiPSC相关的不同分子反应， CM到KIs。我们将进行标准的表型分析、表达分析和功能分析。 用于心脏毒性的测定。随后我们将进行途径表达分析以识别 与KI心脏毒性相关的新功能网络。此外，我们将利用 先前获得的GWAS和全外显子组测序数据以进行表达 定量性状基因座分析，以确定与KI诱导的心脏毒性相关的变异。 我们的建议适用于精确和系统医学的方法来研究心脏毒性。 此外，所提出的方法可以为未来的发展提供重要的见解 新的KIs具有降低的心脏毒性风险特征。

Abstract The development of kinase inhibitors (KIs) has revolutionized the treatment of a broad spectrum of cancer types. However drug-related adverse effects often counter the benefits of anti-cancer drugs. With the widespread use of KIs, it has become apparent that a subset of patients develops cardiac dysfunction and heart failure. Currently our knowledge of the underlying pathophysiology and risk factors is limited. Experiments focusing on understanding the molecular and genetic mechanisms underlying cardiotoxicity will be critical to improve and guide further drug development. The recent development of human induced pluripotent stem cells (hiPSCs) and the ability to differentiate hiPSCs into CMs (hiPSC-CMs) offer unprecedented opportunities for disease modeling and cardiotoxicity testing. Human iPSC-CMs exhibit properties highly similar to their primary counterparts, thus providing a relevant `patient in a dish' model. The overarching goal of this proposal is to identify and understand the molecular and genetic mechanisms underlying KI-induced cardiotoxicity by using hiPSC-CMs. The proposal builds on extensive infrastructure funded through various NHLBI grants. We have generated 250 hiPSCs lines and differentiated these lines into CMs from participants in the NHLBI HyperGEN cohort. We propose to use these hiPSC-CMs to identify underlying pathways and genetic markers, which contribute to the variability in the response to KIs. Using expression analysis, we will describe the distinct molecular responses associated with exposing hiPSC- CMs to KIs. We will perform standard phenotypic analyses, expression analysis and functional assays for cardiotoxicity. Subsequently we will perform pathway expression analysis to identify novel functional networks associated with KI cardiotoxicity. In addition, we will be utilizing previously obtained GWAS and whole exome sequencing data to perform expression quantitative trait locus analysis to determine variants associated with KI-induced cardiotoxicity. Our proposal applies a precision and systems medicine approach to the study of cardiotoxicity. Furthermore the proposed approach can provide important insights for the future development of novel KIs with a reduced cardiotoxic risk profile.